Purification Of Lutein Research Articles

Isolation, purification and cytotoxic evaluation of lutein from mixotrophically grown Chlorella sorokiniana TH01

Mixotrophic cultivation of several microalgae simultaneously utilizing two energy sources of light and organic carbon has been reported to significantly achieve higher biomass productivity than phototrophic and heterotrophic modes. In addition, microalgae have been reported as a promising alternative source of marigold for lutein production. In this work, C. sorokiniana TH01 strain was mixotrophically grown on glucose for production of biomass and lutein. Results revealed that algal biomass productivity, lutein content and productivity reached 0.86–2.08 g/L/d, 4.62–6.89 mg/g and 3.98–14.35 mg/L/d, respectively, from 10 g/L glucose, 3 g/L NaNO3 and 2% (v/v) CO2, under light intensity of 50–500 μmol/m2/s and temperature of 27 °C for 10 days of the cultivation. The purification of lutein from the algal biomass was completed by a novel process of simultaneous cell disruption and extraction and silica-based column chromatography purification, reaching lutein purity of 98%. Moreover, the algal lutein can be preserved by solvation with 0.05 wt% ascorbic acid under temperature of <4 °C in the darkness, achieving 90% stability for 120 days of the storage. Chemical structure of the purified algal lutein was fully characterized by NMR and UV–vis spectroscopy and MS spectrometry, showing similar characteristics as the standard lutein and other lutein reported in the literature. Furthermore, the analysis of antiproliferative activity of the algal lutein against human KB, HepG2, MCF-7 and SK-LU-1 cancer cell lines using SRB assay revealed that the algal lutein exhibited moderate activities with corresponding IC50 values determined of 58.07, 61.37, 71.52 and 81.69 μg/mL, respectively. Despite unexpected cytotoxicity on these targeted human cancer cell lines, the algal lutein is a high value compound for commercial applications in food, feed, pharmaceutical and cosmetic industries. The C. sorokiniana TH01 strain was demonstrated to be a promising lutein source, alternatively replacing marigold-based lutein.

Recent advances in lutein production from microalgae

Lutein is a xanthophyll carotenoid with remarkable applications in the healthcare sector due to its ocular-protective, anti-oxidative and anti-inflammatory activities. Currently, marigold flower petals are the only commercial source for lutein extraction, since chemical synthesis suffers from very low yields. Microalgae cultivation is a promising way for lutein production, as the lutein content of microbial biomass is higher compared to that of marigold petals. In addition, microalgae are shown to have a higher growth rate, higher photosynthetic efficiency, and carbon mitigation potential when compared to terrestrial plants, such as marigolds. For the realization of microalgae as a sustainable and alternative lutein source, crucial factors such as the microalgal strains, critical cultivation parameters (light intensity, nutritional status, metabolic mode), pretreatment, extraction, and purification of lutein from microalgal biomass need to be studied in great detail. In this context, this review summarized the recent research progress in lutein production from microalgae, including competent microalgal strains, cultivation systems, and subsequent biomass treatment technologies. The choice of a high yielding strain, along with effective photobioreactor design will enable maximal lutein production from microalgae in an economically viable manner. This review presents comprehensive information on microalgal cultivation and pretreatment which will enable researchers to design an optimal microalgae-based lutein production system.

Purification of lutein from the green microalgae Chlorella vulgaris by integrated use of a new extraction protocol and a multi-injection high performance counter-current chromatography (HPCCC)

Lutein is a natural carotenoid with health promoting properties whose commercial source is the yellow Marigold flower, but its production is limited by climate, planting area and high labor costs. The microalga Chlorella vulgaris is a promising lutein source; however, no commercial lutein production uses microalgae. In the present study, the lower phase (LP4) of the biphasic solvent system composed of n-heptane–ethanol–water (5:4:1.5, v/v/v) was used both as a solvent for microalgae biomass extraction and as a mobile phase for lutein isolation using high performance counter-current chromatography (HPCCC). The ultrasound-assisted extraction of biomass with LP4 for 30 min led to an extract with a lutein content (3.20 mg/g dried biomass) that was higher than those obtained by the reference solvents tetrahydrofuran and dichloromethane. A multi-injection HPCCC method was developed for lutein isolation, where LP4 was pumped as the mobile phase at a flow rate of 8 mL/min, whereas its upper phase was the stationary phase. In total, 2 g of C. vulgaris extract were processed affording 60 mg of lutein (92% purity), which was further cleaned up by gel permeation chromatography yielding 50 mg of lutein (97% purity). The chemical identity of the purified compound was confirmed by mass spectrometry and absorption spectroscopy in comparison with an authentic standard. The developed HPCCC method may serve as a reference for promoting the development of a new production model of microalgae-based lutein.

Quantification and purification of lutein and zeaxanthin recovered from distillers dried grains with solubles (DDGS)

Marigold petals, the current commercial source for lutein and zeaxanthin are harvested through a labor-intensive operation with downstream purification requiring multiple processing steps involving various harsh solvents. Lutein and zeaxanthin are in-demand carotenoids due to their significant role in human eye health. A possible alternative source for lutein and zeaxanthin, distillers dried grain with solubles (DDGS), shows promise and contains a yet-to-be quantified amount of the desired carotenoids. The US corn industry produces an abundant, and in some cases excess, annual supply of DDGS. The large volume of DDGS produced could serve as a significant source for lutein and zeaxanthin recovery and provides an additional market stream for the growing carotenoid industry. This paper demonstrates one of the first quantitative reports regarding the concentration of lutein and zeaxanthin in DDGS. Using Soxhlet extraction, followed by purification with centrifugal partition chromatography, it was determined that 36.09 ± 16.87 µg lutein and 15.48 ± 6.13 µg zeaxanthin could be purified from all extractives retained in the oleoresin per gram of DDGS. As compared to lutein and zeaxanthin present in corn, this is a three to fivefold increase indicating that these compounds become concentrated during the dry grind process. Recovery of lutein and zeaxanthin from DDGS, a low-value stream by-product of corn ethanol industry, results in a new revenue stream and would add value to a common US commodity.

Combined process of reaction, extraction, and purification of lutein in marigold flower by isopropanol–KOH aqueous two-phase system

ABSTRACTThis paper established a viable process for the simultaneous reaction, extraction, and primary purification of lutein in marigold flower by the isopropanol–KOH aqueous two-phase system (ATPS). In this process, the liposoluble lutein esters were translated into water-soluble lutein by saponification reaction in ATPS and distributed to the top phase. Meanwhile, the water-soluble flavonoids in marigold were also extracted and distributed to the bottom phase. The concentrations of phase-forming isopropanol and KOH, the added mass of marigold flower powder, temperature, and time were investigated to evaluate their effects on the extraction and partition behaviour of lutein and flavoniods in ATPS. Under the optimal conditions (38% isopropanol, 18% KOH, 0.05 g of marigold flower powder, 500°C and 1.5 h extraction time), the leaching efficiency, extraction efficiency, and partition coefficient of lutein were 84.64%, 86.73%, and 5.115, respectively, and at this point, the saponification rate of lutein reached 93.98%. Meanwhile, the leaching efficiency, extraction efficiency, and partition coefficient of flavoniods were 96.68%, 78.31%, and 0.2167, respectively. This method shows the superiority to the conventional one for it can not only efficiently extract lutein in marigold flower, but also achieve a high saponification rate of lutein esters and simultaneously separate two major pigments.

Isolation and Purification of Lutein from Indian SpinachBasella alba

Lutein is a pharmacologically important phytochemical belonging to the class of carotenoids widely distributed in marigold flowers and green vegetables. It is industrially important for its biological activities such as antioxidant, immunostimulatory and macular protective properties. In this study, isolation and purification of lutein from Indian spinach, Basella alba was carried out. For the extraction of lutein, petroleum ether-acetone extract of spinach leaves was prepared which yielded 2.3 mg/100g of lutein. Preparative TLC of the extract yielded lutein with a Rf value of 0.12. Lutein isolated was subjected to spectral characterization by UV-Visible and FT-IR spectroscopy. The peaks obtained corresponded well with those reported for standard lutein. Results of the study indicated that B. alba leaves can be used as a source for obtaining lutein.

Recovery of Free Lutein From Tagetes erecta: Determination of Suitable Saponification and Crystallization Conditions

This study deals with the separation and purification of lutein from marigold flowers. The effects of saponification conditions such as time (0–7 hr), volume to weight ratio of KOH solution to oleoresin (0.3 ml:1 g to 1 ml:1 g), and the concentration of KOH solution (25%–45% w/v) were first determined on the conversion of lutein fatty acid esters to free lutein. For 1 g of oleoresin, 0.5 ml of 40% v/v KOH solution was found to be the most suitable amount and concentration for the saponification of lutein esters to free lutein at 75°C. Following saponification, crystallization was carried out to determine the effect of crystallization solvents (water-ethanol and water-isopropanol mixtures), the composition of the crystallization solvents (2:0.3–2:1 v/v), and crystallization time (0.25–2 hr) on the yield and purity of free lutein. The results revealed that crystallization of the saponified sample in water-ethanol mixture at the volume ratio of 2:0.5 for 0.5 hr presented the highest yield of 99.12% of free lutein, at the purity of 85.53%.

Isolation and purification of lutein from the microalga Chlorella vulgaris by extraction after saponification.

A simple and efficient method for the isolation and purification of lutein from the microalga Chlorella vulgaris was developed. Crude lutein was obtained by extraction with dichloromethane from the microalga after saponification. Partition values of lutein in the two-phase system of ethanol-water-dichloromethane at different ratios were measured by HPLC so as to assist the determination of an appropriate condition for washing water-soluble impurities in the crude lutein. Partition values of lutein in another two-phase system of ethanol-water-hexane at different ratios were also measured by HPLC for determining the condition for removing fat-soluble impurities. The water-soluble impurities in the crude lutein were removed by washing with 30% aqueous ethanol, and the fat-soluble impurities were removed by extraction with hexane. The final purity of lutein obtained was 90-98%, and the yield was 85-91%.

Preparative isolation and purification of lutein from the microalga Chlorella vulgaris by high-speed counter-current chromatography

High-speed counter-current chromatography (HSCCC) was applied to the isolation and purification of lutein from microalgae. Analytical HSCCC was used for the preliminary selection of a suitable solvent system composed of n-hexane–ethanol–water (4:3:1, v/v). Using the above solvent system, preparative HSCCC was successfully performed yielding lutein at 98% purity from 200 mg of the crude extract in a one-step separation.